Quantitative analysis of oligonucleotides by matrix-assisted laser desorption/ionization mass spectrometry

Rapid Communications in Mass Spectrometry : RCM
B A BruennerT W Hutchens

Abstract

Quantitative aspects of oligonucleotide analysis by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry remain largely unexplored relative to the efforts that have been devoted to quantitative peptide and protein analysis. The successful quantitation of these other biopolymers coupled with the potential of rapid nucleic acid analysis by desorption/ionization techniques prompted the present investigation into quantifying mixed base oligonucleotides of intermediate molecular weights. This report describes the concentration-dependent desorption/ionization of a 21-base oligonucleotide (MW 6361) using a 36-base oligonucleotide (MW 11 131) as an internal standard. Peak height and peak area ratios (analyte to internal standard) varied linearly as a function of oligonucleotide concentration (R2 = 0.966 and 0.991, respectively). The linearity of response extended over nearly three orders of magnitude, from 0.125 to 100 pmol of analyte applied. The use of an internal standard improved the linearity of the calibration curve and reduced relative standard deviations. These results demonstrate for the first time the quantitation of medium size oligonucleotides using MALDI.

Citations

Nov 29, 2005·Clinical Chemistry and Laboratory Medicine : CCLM·Roisin M MolloyStephen P FitzGerald
Nov 5, 1999·Analytical Biochemistry·C E MacPheeW H Sawyer
Apr 1, 2003·Mass Spectrometry Reviews·Jörg Tost, Ivo G Gut
Feb 8, 2005·Molecular & Cellular Proteomics : MCP·Michael E WrightRuedi Aebersold
Apr 23, 2008·Current Protocols in Nucleic Acid Chemistry·C W Chou, P A Limbach
Jul 27, 2001·Journal of Immunological Methods·R P Huang
Sep 27, 2002·Journal of the American Society for Mass Spectrometry·Martin BucknallMark W Duncan
Dec 14, 2004·Journal of the American Society for Mass Spectrometry·Ying L Li, Michael L Gross

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